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Nat Methods. 2018 Aug;15(8):617-622. doi: 10.1038/s41592-018-0044-9. Epub 2018 Jul 9.

Genome-wide SWAp-Tag yeast libraries for proteome exploration.

Author information

1
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
2
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
3
Département de Biochimie, Faculté de Médecine, Université de Montréal, Montreal, QC, Canada.
4
Department of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot, Israel.
5
Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.
6
Department of Microbiology and Molecular Genetics, IMRIC, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel.
7
Department of Cell Biology, University of Alberta, Edmonton, AB, Canada.
8
Department of Cell Biology, University of Kaiserslautern, Kaiserslautern, Germany.
9
Department of Life Sciences Core Facilities, Weizmann Institute of Science, Rehovot, Israel.
10
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel. maya.schuldiner@weizmann.ac.il.

Abstract

Yeast libraries revolutionized the systematic study of cell biology. To extensively increase the number of such libraries, we used our previously devised SWAp-Tag (SWAT) approach to construct a genome-wide library of ~5,500 strains carrying the SWAT NOP1promoter-GFP module at the N terminus of proteins. In addition, we created six diverse libraries that restored the native regulation, created an overexpression library with a Cherry tag, or enabled protein complementation assays from two fragments of an enzyme or fluorophore. We developed methods utilizing these SWAT collections to systematically characterize the yeast proteome for protein abundance, localization, topology, and interactions.

PMID:
29988094
PMCID:
PMC6076999
DOI:
10.1038/s41592-018-0044-9
[Indexed for MEDLINE]
Free PMC Article

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